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Journal of Virology, June 2002, p. 5882-5892, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.5882-5892.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Development and Characterization of Bovine x Human Hybrid Cell Lines That Efficiently Support the Replication of both Wild-Type Bovine and Human Adenoviruses and Those with E1 Deleted

Alberto L. van Olphen,{dagger} and Suresh K. Mittal*

Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, Indiana 47907

Received 4 January 2002/ Accepted 12 March 2002

The 293 cell line that was generated by transforming human embryonic kidney cells with human adenovirus type 5 (HAV5) early region 1 (E1) sequences is an excellent host for generating and growing HAV5 recombinants with E1 deleted, but it does not support the replication of bovine adenovirus type 3 (BAV3). Madin-Darby bovine kidney (MDBK), an established bovine cell line, is an excellent host for growing and plaquing BAV3. For the purpose of combining the unique characteristics of these two cell lines (293 and MDBK), we generated a number of bovine x human hybrid (BHH) cell lines. Comparison of three BHH hybrid clones—BHH3, BHH8, and BHH2C—with 293-Puro (puromycin-resistant 293 cells) and MDBK-Neo (G418-resistant MDBK cells) cell lines for total cellular DNA content, species-specific surface markers, isoenzyme analysis, and karyotyping indicate that they are hybrid in nature. BHH clones constitutively expressed the E1 proteins (E1A, E1B-21kDa, and E1B-55kDa) of HAV5 and efficiently supported the replication of both wild-type and replication-incompetent bovine or human adenoviruses. Transient gene expression experiments with a plasmid encoding the bacterial ß-galactosidase gene demonstrated that BHH cell hybrids seem to have better transfection efficiencies than either of the parental cell lines. These cell lines will be useful for isolating and growing replication-competent human or bovine adenovirus recombinants with E1 deleted and for the study of cellular or viral factors important for viral replication. The development of somatic cell hybrids appears to be a simple way of combining some of the desirable characteristics present separately in two parental cell lines.

* Corresponding author. Mailing address: Department of Veterinary Pathobiology, 1290 Lynn Hall, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907. Phone: (765) 496-2894. Fax: (765) 494-9830. E-mail: mittal{at}purdue.edu.

{dagger} Present address: Wyoming State Veterinary Laboratory, Department of Veterinary Science, University of Wyoming, Laramie, WY 82070.

Journal of Virology, June 2002, p. 5882-5892, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.5882-5892.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Ali, S., Pellerin, C., Lamarre, D., Kukolj, G. (2004). Hepatitis C Virus Subgenomic Replicons in the Human Embryonic Kidney 293 Cell Line. J. Virol. 78: 491-501 [Abstract] [Full Text]  


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